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1.
Methods Mol Biol ; 2144: 29-45, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32410022

RESUMEN

We miniaturized standard, solid-phase C. elegans culture conditions to produce a system in which many isolated, individual C. elegans can be housed throughout their lives. This system, the "worm corral," is compatible with high-resolution brightfield and fluorescent microscopy, allowing imaging of fluorescent transgenes and morphological phenotypes from hatch until death. These culture devices can be constructed on the benchtop with commercially available reagents and standard laboratory equipment, making this an attainable solution for most labs.


Asunto(s)
Caenorhabditis elegans/crecimiento & desarrollo , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/métodos , Animales , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Microscopía Fluorescente , Fenotipo
2.
Lab Chip ; 17(22): 3909-3920, 2017 11 07.
Artículo en Inglés | MEDLINE | ID: mdl-29063084

RESUMEN

We have miniaturized standard culture techniques to rear arrays of isolated, individual C. elegans throughout their lives on solid gel media. The resulting apparatus is compatible with brightfield and fluorescence microscopy, enabling longitudinal studies of morphology and fluorescent transgene expression. Our culture system exploits a novel crosslinking reaction between a polyethylene glycol hydrogel and a silicone elastomer to constrain animals to individual "corrals" on the gel surface. These devices are simple to construct on the benchtop with commercially available reagents, and, unlike microfluidic isolation methods, do not rely on micropatterned materials. We demonstrate that this new culture method has negligible effects on the physiology of C. elegans compared to standard culture on agar plates. In addition, RNAi techniques are effective in this system. Finally, the hydrogel-silicone binding chemistry that we developed also allows traditional microfluidic devices to be covalently attached to gel substrates instead of glass.


Asunto(s)
Caenorhabditis elegans/fisiología , Técnicas de Cultivo/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Microscopía Fluorescente/instrumentación , Animales , Técnicas de Cultivo/métodos , Diseño de Equipo , Hidrogeles , Dispositivos Laboratorio en un Chip , Microscopía Fluorescente/métodos , Polietilenglicoles
3.
Cell Syst ; 3(4): 333-345.e4, 2016 10 26.
Artículo en Inglés | MEDLINE | ID: mdl-27720632

RESUMEN

Although many genetic factors and lifestyle interventions are known to affect the mean lifespan of animal populations, the physiological variation displayed by individuals across their lifespans remains largely uncharacterized. Here, we use a custom culture apparatus to continuously monitor five aspects of aging physiology across hundreds of isolated Caenorhabditis elegans individuals kept in a constant environment from hatching until death. Aggregating these measurements into an overall estimate of senescence, we find two chief differences between longer- and shorter-lived individuals. First, though long- and short-lived individuals are physiologically equivalent in early adulthood, longer-lived individuals experience a lower rate of physiological decline throughout life. Second, and counter-intuitively, long-lived individuals have a disproportionately extended "twilight" period of low physiological function. While longer-lived individuals experience more overall days of good health, their proportion of good to bad health, and thus their average quality of life, is systematically lower than that of shorter-lived individuals. We conclude that, within a homogeneous population reared under constant conditions, the period of early-life good health is comparatively uniform, and the most plastic period in the aging process is end-of-life senescence.


Asunto(s)
Caenorhabditis elegans , Animales , Calidad de Vida
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